Glutathione depletion modulates gene expression in HepG2 cells via activation of protein kinase C alpha

Angela T White, Fiona J Spence, James K Chipman

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)


Buthionine sulphoximine (BSO; 1mM) resulted in the depletion of glutathione (GSH) in HepG2 cells to 17+/-1.5% within 24h. This was not associated with apoptotic or necrotic cell death over this time period. Use of a human (Phase 1) cDNA custom toxicology-array and a larger scale (>10,000 gene) Affymetrix U95Av2 array identified a total of 48 and 104 genes, respectively, with a statistically significant (and >1.5-fold) change in expression. A total of 64 differentially expressed genes (6 of which were confirmed by real-time polymerase chain reaction) were suggestive of protein kinase C (PKC) activation. Activation of PKC-alpha (but not betaI or delta) was demonstrated at 24 h through activity measurements and through Western blot analysis of membrane-associated PKC-alpha protein. Activation did not occur in the presence of additional gamma-glutamylcysteine to prevent GSH depletion. Activation of PKC-alpha by GSH-depletion may, at least in part, be mediated by thiol oxidation and may contribute to a survival signal. If sustained, the activation may be important in non-genotoxic carcinogenesis.
Original languageEnglish
Pages (from-to)168-80
Number of pages13
Issue number2-3
Publication statusPublished - 15 Dec 2005


  • Animals
  • Apoptosis
  • Oligonucleotide Array Sequence Analysis
  • Protein Kinase C-alpha
  • Carcinoma, Hepatocellular
  • Enzyme Activation
  • Glutathione
  • Humans
  • Buthionine Sulfoximine
  • Cell Line, Tumor
  • Reverse Transcriptase Polymerase Chain Reaction
  • Protein Isoforms
  • Cell Survival
  • Liver Neoplasms
  • L-Lactate Dehydrogenase
  • Gene Expression Profiling
  • Blotting, Western
  • Gene Expression Regulation, Enzymologic
  • Phosphorylation
  • Protein Kinase C
  • Time Factors
  • Protein Transport


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